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Biomedical subjects

G Blaschke

Publications and source records attributed to G Blaschke.

At least 55 records · Page 3Linked to original sources
PubMed1999

Achiral and chiral determination of ciprofibrate and its glucuronide in human urine by capillary electrophoresis.

A method for achiral separation of the racemic hypolipidaemic agent ciprofibrate and its main metabolite ciprofibrate glucuronide by capillary electrophoresis (CE) was developed. The glucuronide was isolated from urine by chromatographic procedures and characterized by alkaline as well as enzymatic hydrolysis and mass spectrometric and nuclear magnetic resonance experiments. Chiral discrimination of the ciprofibrate enantiomers and their diastereomeric glucuronides by CE was achieved by the use of gamma-cyclodextrin as buffer additive. The fractionated crystallization of ciprofibrate yielded the R-(+)-enantiomer as less soluble diastereomeric salt with (+)-1-phenylethylamine. This allowed the identification of the enantiomers of ciprofibrate as well as the diastereomeric glucuronides of ciprofibrate by CE. In a study with three volunteers inter- and intra-individual differences of ratios of both ciprofibrate glucuronides in urine were observed. After oral administration of a single dose of the racemate the S-ciprofibrate glucuronide was mainly excreted in the first time intervals, in the last time intervals the R-glucuronide dominated.

Clofibric Acid↗
PubMed1999

Determination of (E)-5-(2-bromovinyl)-2'-deoxyuridine in plasma and urine by capillary electrophoresis.

(E)-5-(2-Bromovinyl)-2'-deoxyuridine is an antiviral drug used for treatment of infections with Herpes simplex virus type 1 as well as Varicella zoster virus. Two fast methods for the determination of the drug and its metabolite in plasma and urine by capillary electrophoresis have been developed. The plasma method can be used for measurement of total as well as unbound drug and metabolite. Plasma and urine samples are prepared for measuring by liquid/liquid extraction resulting in a limit of quantification of 40 ng/ml for total and 10 ng/ml for free BVdU in plasma and 170 ng/ml in urine. Inter- as well as intra-day precision were found to be better than 10% and both methods have been used for drug monitoring of patients.

Antiviral Agents↗
PubMed1999

Immunoaffinity extraction of morphine, morphine-3-glucuronide and morphine-6-glucuronide from blood of heroin victims for simultaneous high-performance liquid chromatographic determination.

The development of an immunoaffinity-based extraction method for the determination of morphine and its glucuronides in human blood is described. For the preparation of an immunoadsorber, specific antisera (polyclonal, host: rabbit) against morphine, morphine-3-glucuronide and morphine-6-glucuronide were coupled to 1,1'-carbonyldiimidazole-activated tris-acrylgel and used for immunoaffinity extraction of morphine and its glucuronides from coronary blood. The resulting extracts were analysed by HPLC with native fluorescence detection. The mean recoveries from spiked blood samples were 71%, 76% and 88% for morphine, morphine-3-glucuronide and morphine-6-glucuronide, respectively. The limit of detection was 3 ng/g blood and the limit of quantitation was 10 ng/g blood for all three analytes. The results of the analysis of coronary blood samples from 23 fatalities due to heroin are presented.

Chromatography, Affinity↗
PubMed1999

Investigation of the in vitro biotransformation of R-(+)-thalidomide by HPLC, nano-HPLC, CEC and HPLC--APCI-MS.

High-performance liquid chromatography (HPLC), nano-HPLC, capillary electrochromatography (CEC) and on-line HPLC-atmospheric pressure chemical ionization mass spectrometry (APCI-MS) techniques were used for the identification and detailed characterization of two new metabolites of the former sedative drug thalidomide (TD). The advantages of nano-HPLC and CEC are higher peak efficiency and a drastic decrease in the analysis time, which, together with lower sample dilution during the analyses, allowed to obtain a detection sensitivity that was comparable to HPLC with common-sized columns. Both, nano-HPLC and CEC could be realized in the commercially available capillary electrophoresis system HP3D. On-line HPLC-APCI-MS coupling is a very useful technique for the rapid identification of metabolites without any need for reference compounds.

Animals↗
PubMed1999

Direct determination of paracetamol and its metabolites in urine and serum by capillary electrophoresis with ultraviolet and mass spectrometric detection.

The use of capillary electrophoresis (CE) for the determination of paracetamol and its main metabolites in urine and serum is described. Due to its high efficacy, CE enables the analysis of drugs directly in complex matrices. Thus, simple, rapid and reliable assays could be developed that made use of some of the main advantages of this analytical technique. In order to prevent the peaks from tailing, a water zone was injected behind the sample. Occasionally occurring peak splittings of paracetamol were investigated and methods to suppress these splittings were developed. Paracetamol, its main metabolites, paracetamol glucuronide, paracetamol sulfate as well as paracetamol cysteinate and paracetamol mercapturate, as metabolites of the oxidative pathway were identified in urine using diode-array detection and coupling of the CE instruments to electrospray-mass spectrometry. The assays were validated. Their usefulness was demonstrated by applying them to the analysis of urine and serum samples of healthy volunteers as well as to urine samples from children under anticancer therapy.

Acetaminophen↗
PubMed1999

Direct assay of nonopioid analgesics and their metabolites in human urine by capillary electrophoresis and capillary electrophoresis-mass spectrometry.

Capillary electrophoresis (CE) was used for the analysis of nonopioid analgesics and their metabolites directly in urine samples. A simple, reliable screening method was developed that allows identification of the drug and/or its metabolites in urine after oral application of paracetamol, acetylsalicylic acid, antipyrine, ibuprofen, naproxen, ketoprofen and propyphenazone by their migration in CE and by their UV spectra recorded with a diode-array detector in a common CE-UV system with 50 mM borax pH 9.4 as separation buffer. For the CE-electrospray (ESI)-MS coupling a volatile 50 mM ammonium acetate buffer at pH 9.8 was used. In order to analyze the metabolic pattern in more detail different methods were developed for each drug. The separation of the metabolites of acetylsalicylic acid could be improved by injection of the urine sample at the cathodic side of the capillary. In order to identify antipyrine as neutral compound as well as its neutral metabolites-a micellar electrokinetic chromatography (MEKC) method was developed.

Analgesics, Non-Narcotic↗
PubMed1999

Enantioseparation of amfepramone (rac-diethylpropion): preparative separation of the enantiomers and enantioselective analysis.

The enantiomers of the anorectic drug amfepramone [rac-diethylpropion, rac-2-(diethylamino)-1-phenyl-1-propanone; rac-DEP] were separated in the preparative scale by crystallization. With enantiopure di-O-benzoyltartaric acid as salt-forming chiral selector, diastereoisomeric salts of DEP enantiomers with a final purity of more than 97.5% were obtained. Analytical liquid chromatographic and electrophoretic methods for the control of the enantiomeric purity and the stoichiometry of the salts were developed. The enantioseparation of rac-DEP by capillary electrophoresis (CE) using hydroxypropyl-beta-cyclodextrin (HP-beta-CD) as chiral discriminator and phosphate buffer (pH 3.3) as run buffer led to good separations. HPLC methods were developed using polysaccharide chiral stationary phases (CSP). The separation of the two enantiomers and the two main degradation products (1-phenyl-1,2-propanedione and propiophenone), known from solid and liquid pharmaceutical preparations, was attained in one run on the silica-based CSP cellulose tris(3,5-dimethylphenylcarbamate) (Chiralcel OD). The conditions which might affect the enantioselectivity and the quality of the enantiomeric separation were investigated for Chiralcel OD and the related CSP amylose tris(3,5-dimethylphenylcarbamate) (Chiralpak AD). Both CSPs showed very similar chromatographic properties. The separation factors could be influenced significantly by varying the polar organic modifier added to the mobile phase.

Appetite Depressants↗
PubMed1999

Chiral recognition of verapamil by cyclodextrins studied with capillary electrophoresis, NMR spectroscopy, and electrospray ionization mass spectrometry.

Capillary electrophoresis (CE) allows the observation of the opposite affinities of the enantiomers of (+/-)-verapamil [2-isopropyl-2,8-bis(3,4-dimethoxyphenyl)-6-methyl-6-azaoctannitrile+ ++, VP] toward beta-cyclodextrin (beta-CD) and heptakis(2,3, 6-tri-O-methyl)-beta-CD (TM-beta-CD). In addition, in the presence of beta-CD in the background electrolyte, longer migration times and lower separation factors were observed compared to TM-beta-CD. The binding constants of (+)- and (-)-VP with beta-CD and TM-beta-CD determined using (13)C NMR spectroscopy explain the results observed in CE. Electrospray ionization mass spectrometry (ESI-MS) was used as an alternative technique for the characterization of VP-CD complexes.

Calcium Channel Blockers↗
PubMed1999

Achiral and chiral determination of tramadol and its metabolites in urine by capillary electrophoresis.

An achiral and chiral separation for the determination of tramadol and its main metabolite O-demethyltramadol in urine samples by CE with UV detection was developed. It was possible to separate tramadol and its phase I and phase II metabolites in one single run using a borate buffer. Furthermore, the simultaneous chiral separation of tramadol and the phase I metabolites was achieved using carboxymethyl-beta-cyclodextrin as chiral selector. To reach the required limits of quantification for the analytes, a preconcentration by solid-phase extraction for the achiral assay and by liquid-liquid extraction for the chiral assay was used. The methods were validated and their applicability was shown by the determination of tramadol and O-demethyltramadol in urine samples.

Analgesics, Opioid↗
PubMed1999

Enantioseparation of thalidomide and its hydroxylated metabolites using capillary electrophoresis with various cyclodextrins and their combinations as chiral buffer additives.

The separation of thalidomide (TD) and its hydroxylated metabolites including their simultaneous enantioseparation was studied in capillary electrophoresis (CE) using four different randomly substituted charged cyclodextrin (CD) derivatives, the combinations of some of them with each other, and beta-CD. TD, as well as two metabolites recently found in incubations of human liver microsomes and human blood, 5-hydroxythalidomide (5-OH-TD) and one of the diastereomeric 5'-hydroxythalidomides (5'-OH-TD), are neutral compounds. Therefore, they were resolved using charged chiral selectors in CE. Two different separation modes (normal polarity and carrier mode) and two different capillaries (fused-silica and polyacrylamide-coated) were tested. Based on the behavior of the individual CDs, their designed combinations were selected in order to improve the separation selectivity and enantioselectivity. Under optimized conditions all three chiral compounds and their enantiomers were resolved simultaneously.

Acrylic Resins↗
PubMed1999

Selector-selectand interactions in chiral capillary electrophoresis.

This review discusses selected aspects of selector-select and interactions in chiral capillary electrophoresis (CE). Studies performed using nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS) and X-ray crystallography for a better understanding of chiral recognition mechanisms in CE are summarized. The theoretical background of chiral CE in general, mathematical models, method development and optimization strategies, etc., are not covered. A general overview on the most recent developments in chiral CE is presented in this volume in the review paper by Bocek [1].

Crystallography, X-Ray↗
PubMed1999

Potential of flow-counterbalanced capillary electrophoresis for analytical and micropreparative separations.

The potential of flow-counterbalanced capillary electrophoresis (FCCE) in chiral and achiral separations was investigated in this work. Unlimited increase of the separation selectivity can be achieved for binary mixtures using FCCE. This was shown for the enantioseparation of (+/-)-chlorpheniramine (CHL) with carboxymethyl-beta-cyclodextrin (CM-beta-CD) as chiral selector. The other example is the separation of alpha- and beta-isomers of a dipeptide aspartame (AS). The carrier ability of the (chiral) selector or pseudostationary phase, the electroosmotic flow (EOF), the pressure-driven flow or hydrodynamic flow can be used as a counterbalancing flow to the electrophoretic mobility of the analyte or vice versa. This mechanism can also be used for micropreparative purposes. FCCE also bears the potential for stepwise separation and fraction collection of multicomponent mixtures.

Electrophoresis, Capillary↗
PubMed1999

Designed combination of chiral selectors for adjustment of enantioseparation selectivity in capillary electrophoresis.

In this study an attempt has been made to explain the reasons for changing the enantioseparation selectivity in some dual cyclodextrin (CD) systems compared to the use of single chiral selectors in capillary electrophoresis (CE). An explanation for selectivity changes is proposed based on the effect of the chiral selector on the mobility of the analyte. In order to support the proposed mechanism, several dual systems were designed on the basis of the known recognition pattern of enantiomers for individual CDs. In most cases the separation selectivity could be adjusted in a designed way. There was no experimental evidence for simultaneous binding of a given chiral analyte with both chiral selectors or of chiral recognition of an analyte complex with one CD by another CD.

Electrophoresis, Capillary↗
PubMed1999

Chiral separations in capillary high-performance liquid chromatography and nonaqueous capillary electrochromatography using helically chiral poly(diphenyl-2-pyridylmethyl methacrylate) as chiral stationary phase.

Enantioseparations in nonaqueous capillary electrochromatography (CEC) are reported in this study for the first time, using wide-pore aminopropyl silica gel coated with helically chiral poly(diphenyl-2-pyridylmethyl methacrylate) (PDPM) as chiral stationary phase (CSP). The anodic electroosmotic flow (EOF) in a methanolic solution of ammonium acetate was used for the migration of neutral analytes through the packed bed in the capillaries. Four different techniques, high-performance liquid chromatography (HPLC) in common-size columns, capillary HPLC, pressure-assisted CEC and CEC were compared from the viewpoint of separation parameters. The latter three were performed with the same experimental setup, varying the relative contribution of the pressure-driven and the electrically driven flow to the overall mobility of the analyte. Capillary HPLC offers clear advantages compared to enantioseparations in common-size columns. However, for a given particle size of the packing material, CEC was not obviously advantageous compared to pressure-driven separations.

Chromatography, High Pressure Liquid↗
PubMed1999

Investigation of the major human hepatic cytochrome P450 involved in 4-hydroxylation and N-dechloroethylation of trofosfamide.

Trofosfamide and its congeners ifosfamide and cyclophosphamide are cell-cycle-nonspecific alkylating agents that undergo bioactivation catalyzed by liver cytochrome P450 (CYP) enzymes. Two NADPH-dependent metabolic routes for the anticancer drug trofosfamide, i.e., 4-hydroxylation and N-dechloroethylation, were studied in human liver microsomes and in seven recombinant human CYP isoforms (i.e., CYP1A1, 1A2, 2A6, 2B6, 2D6, 2E1, and 3A4-OR) to identify the CYP enzymes involved. Recombinant human CYP3A4 and CYP2B6 exhibited catalytic activity with respect to both pathways of trofosfamide. Enzyme kinetic analyses revealed the dominant role of human CYP3A4 in 4-hydroxylation and N-dechloroethylation of trofosfamide. This was confirmed by the observation that only the CYP3A4 contents of five samples of human liver microsomes correlated with both pathways of trofosfamide. Furthermore, ketoconazole, a selective inhibitor of CYP3A4, substantially inhibited microsomal trofosfamide 4-hydroxylation and N-dechloroethylation (50% inhibitory concentration < 1 microM for both reactions). The present study indicates that human liver microsomal CYP3A4 preferentially catalyzes the two NADPH- dependent metabolic routes of trofosfamide, which emphasizes the necessity for awareness of potential interactions with any coadministered drugs that are CYP3A4 substrates.

Alkylation↗
PubMed1999

Stereoselectivity of the phase-II-metabolism of the H1 antihistaminic drug dimethindene investigated by capillary electrophoresis.

Two capillary electrophoretic (CE) methods for the determination of conjugated metabolites of the antihistaminic drug dimethindene were developed. These methods allow the determination of the diastereomeric ratios of both Phase II glucuronides, 6-hydroxy dimethindene glucuronide (5) and 6-hydroxy-N-demethyl dimethindene glucuronide (6). Furthermore, after enzymatic deconjugation of both glucuronides, enantiomeric ratios of the hydroxylated Phase I metabolites 3 and 4 were determined. For comparison, glucuronides 5 and 6 were prepared by enzymatic synthesis and identified by MALDI-TOF mass spectrometry and deconjugation with beta-glucuronidase to the corresponding phenols 3 and 4. The assay of the glucuronides in rat urine and serum was performed by a combination of liquid/liquid extraction followed by solid phase extraction. After a high dose treatment with racemic dimethindene the diastereomeric glucuronides 5 and 6 could be directly determined in the urine of a guinea pig by CE without prior extraction or preconcentration of the urine sample. In all cases, stereoselectivities of the formation of Phase I as well as Phase II metabolites were observed.

Animals↗
PubMed1998

Capillary electrophoretic drug monitoring of methotrexate and leucovorin and their metabolites.

High-dose methotrexate is an important element of treatment protocols in childhood acute lymphocytic leukemia or osteosarcoma. A capillary electrophoretic method has been developed to measure peak levels and the metabolite pattern in patients with delayed methotrexate elimination. It serves to determine plasma levels of methotrexate, leucovorin and their metabolites, 7-hydroxymethotrexate, 2,4-diamino-N10 -methylpteroic acid and 5-methyltetrahydrofolic acid. For the determination of high concentrations (>10 microM) protein precipitation by acetonitrile will suffice for sample preparation. All other samples undergo solid-phase extraction and upgrading on C18 columns. Aminopterin, a therapeutic antecedent of methotrexate, serves as internal standard. Detection is done on a UV detector with a 300 nm filter. Five hundred microl of serum are needed to determine 0.2 microM of a specified substance (0.5 microM 5-methylterahydrofolic acid) with good precision and accuracy. For peak levels, 20 microl of capillary serum are sufficient.

Antimetabolites, Antineoplastic↗